Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K39/12—Viral antigens
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K39/12—Viral antigens
  • A61K39/205—Rhabdoviridae, e.g. rabies virus
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
  • A61K2039/525—Virus
  • A61K2039/5254—Virus avirulent or attenuated
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
  • A61K2039/541—Mucosal route
  • A61K2039/542—Mucosal route oral/gastrointestinal
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K39/00—Medicinal preparations containing antigens or antibodies
  • A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
  • A61K2039/552—Veterinary vaccine
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
  • C12N2760/00011—Details
  • C12N2760/20011—Rhabdoviridae
  • C12N2760/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
  • C12N2760/00011—Details
  • C12N2760/20011—Rhabdoviridae
  • C12N2760/20061—Methods of inactivation or attenuation
  • C12N2760/20064—Methods of inactivation or attenuation by serial passage

Definitions

• the aim of the invention is to find a means which is suitable for preventing the spread of rabies by artificial immunization of the fox population and for reducing or eliminating contaminated zones.
• an agent suitable for oral immunization which is characterized by an optionally stabilized vaccine according to the invention which is located in a container according to the invention.
• This agent is fixed in a bait suitable as a vehicle and laid out to eat in the contaminated area.
• the bait is eaten, the container is bitten and chewed, thereby releasing the vaccine in the mouth so that it can be absorbed through the oropharyngeal mucosa.
• this immunization leads to lifelong resistance to rabies viruses in foxes.
• the attenuated new rabies virus strain SAD is used as vaccine virus and is used on BHK 21 .
• the novel vaccine according to the invention is produced by storing cells of the virus strain SAD in a buffered medium at a temperature of 33 to 37 ° C., preferably 33 ° C., and a pH of 6.8 to 7.4, preferably 7.4 , preferably by adding a bicarbonate solution, and inoculated with a cell density of 0.5 to 4 ⁇ 10 5 , preferably 2 ⁇ 10 5 cells per ml of medium, preferably in Roux bottles.
• virus After the formation of a complete cell lawn, it is detached, preferably with trypsin-verse solution, and if necessary diluted in accordance with the increased cell count and sown again.
• the virus is harvested from the cell supernatant after approximately 48 to 72 hours, preferably 48 hours. In compliance with these conditions, virus titres of 10 7.0 to 10 7.5 I are regularly obtained. E ./ml received.
• a cell transfer is dispensed with, a higher virus dose can and must be inoculated initially in order to obtain appropriate virus titers.
• the vaccine is exposed to the environment over the 12 to 72 hour period from when the bait is cast to when it is picked up by the fox. Heat, bait or packaging material can drastically reduce the infectivity of the attenuated virus and thereby cancel out its immunizing effect. Therefore, the vaccine is preferably stabilized so that even at elevated temperatures, e.g. 37 ° C, within e.g. No virus titer loss greater than 90% occurs for 72 hours. Conversely, it is expedient if the living virus used is inactivated spontaneously after a certain time.
• the bowl film can, for example, consist of a rigid PVC film made of plasticizer-free hard PVC, preferably 250 p thick, VC content ⁇ 1 ppm and a cover sheet aluminum film, preferably 40 ⁇ thick, which is thermo-lacquered on the inside (vinyl base 5 to 6 g / m 2 ).
• the agent according to the invention consists of a container according to the invention filled with ml, preferably 1.8 to 2 ml, of vaccine optionally stabilized with egg yolk.
• the minimum dose for the field application is preferably 10 7 tissue culture infectious units.
• the agent is now fixed in a bait suitable as a vehicle so that the vaccine is released in the mouth when the bait is eaten and is absorbed through the oropharyngeal mucosa.
• bait e.g. Chicken heads or minced meat can be used.
• An SAD strain of rabies virus adapted to dog kidney cells is used. This is going to the baby Hamster kidney cell line BHK 21 is adapted by incubating it in Roux bottles in BHK growth medium with the addition of 5% calf serum and 3% fetal calf serum at 33 ° C and a pH between 6.8 to 7.4 and at intervals of 2 to 3 days passaged.
• the cells are inoculated in suspension at a cell density of 0.5 to 2 ⁇ 10 5 cells per ml of medium. After approximately 48 hours, the cell lawn is completely closed and is detached with a trypsin solution of verse and diluted approximately three times in accordance with the increased number of cells and sown again. After a further 48 to 72 hours, the virus is harvested from the cell supernatant. Further results of these tests are summarized in Table 1.
• Virus titration is carried out on cover glass cultures of BHK 21 cells in the Multiwell dish from Costar.
• the Costar plate has 24 depressions with a flat bottom, a diameter of 16 mm and a depth of 18 mm.
• the virus suspended in the BHK medium is mixed with 10% egg yolk as a stabilizer and 2 ml of the product in a push-through pack with a dish film made of a 250 p thick, rigid PVC film made of unplasticized rigid PVC with a VC content of ⁇ 1 ppm, and a cover sheet film made of 40 .mu.m thick, thermally lacquered (vinyl base 5 to 6 g / m 2 ) aluminum film filled and sealed airtight.
• This vaccine virus amount of approx. 10 7 IU per 2 ml, intended for field trials, is fixed in chicken heads and designed as bait in this form.

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• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
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• General Health & Medical Sciences (AREA)
• Immunology (AREA)
• Medicinal Chemistry (AREA)
• Microbiology (AREA)
• Public Health (AREA)
• Zoology (AREA)
• Animal Behavior & Ethology (AREA)
• Pharmacology & Pharmacy (AREA)
• Mycology (AREA)
• Veterinary Medicine (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Organic Chemistry (AREA)
• Epidemiology (AREA)
• Genetics & Genomics (AREA)
• Wood Science & Technology (AREA)
• Biotechnology (AREA)
• Biochemistry (AREA)
• General Engineering & Computer Science (AREA)
• Biomedical Technology (AREA)
• Tropical Medicine & Parasitology (AREA)
• Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

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• 1983
  • 1983-08-02 EP EP83810344A patent/EP0100752A3/fr not_active Withdrawn

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